细胞功能测定

In vivo preclinical assays are required to screen potential agents that target the tumor vasculature. Here, a hollow fibre-based assay for the quantification of neovasculature in the presence or absence of an agent that potentially targets tumor neovasculature is described. The neova ...

Reactive oxygen species (ROS) are constantly produced by skeletal muscle and this production is increased during contractile activity. Understanding the role that ROS play in skeletal muscle requires an understanding of the species of ROS produced, the subcellular site of producti ...

Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteinases associated with extracellular matrix degradation, cellular migration, tissue remodeling, and angiogenesis. The activity of MMPs is regulated by the tissue inhibitors of metalloproteinases (TI ...

Cholesterol-rich microdomains present on the plasma membrane appear to play an important role in spatio-temporal regulation of cell signaling and cell adhesion processes. Compositional heterogeneity of these microdomains and their coalescence during cell–cell interact ...

Dye affinity chromatography is a purification technique offering unique selectivities and high purification factors. Dye ligands may act as substrate analogs, offering affinity interactions with their corresponding enzymes. This chapter describes a dye ligand chromatog ...

Displacement chromatography has several advantages over the nonlinear elution technique, as well as the linear elution mode, such as the recovery of purified components at high concentrations, less tailing during elution, high throughput and high resolution. Displacer affinity ...

Immobilized Metal ion affinity chromatography (IMAC) is a ubiquitous technique in Modern recombinant production and purification. The wide range of expression vectors for the production of histidine-tagged recombinant proteins as well as the variety of stationary supports for ...

The HQ (H = histidine, Q = glutamine) tag is a small fusion tag that can be isolated using immobilized metal affinity columns. HQ-tagged proteins can be expressed and purified from bacterial cells under native and denaturing conditions, mammalian cells, insect cells, wheat germ and rabbit retic ...

Maltose-binding protein (MBP) is a carrier protein for high level recombinant protein and peptide production from either the cytoplasm or periplasm of Escherichia coli. The affinity matrix for purifying MBP-passenger proteins utilizes amylose covalently attached to magnetic b ...

Where an affinity tag has served its purpose, it may become desirable to remove it from the protein of interest. This chapter describes the removal of such fusion partners from the intended protein product by cleavage with site-specific endoproteases. Methods to achieve proteolytic cleav ...

HaloTag™ is a protein fusion tag which was genetically engineered to covalently bind a series of specific synthetic ligands. All ligands carry two groups, the reactive group and the functional/reporter group. The reactive group, the choloroalkane, is the same in all the ligands and is involved ...

The use of affinity tags and especially histidine tags (His-tags) has become widespread in molecular biology for the efficient purification of recombinant proteins. In some cases, the presence of the affinity tag in the recombinant protein is unwanted or may represent a disadvantage for the ...

Synthetic affinity ligands can circumvent the drawbacks of natural immunoglobulin (Ig)-binding proteins by imparting resistance to chemical and biochemical degradation and to in situ sterilization, as well as ease and low cost of production. Protein L (PpL), isolated from Peptostr ...

Large repertoires of peptides displayed on bacteriophage have been extensively used to select for ligand-binding molecules. This is a relatively straightforward process involving several cycles of selection against target molecules, and the resulting ligands can be tailored ...

monolithic macroporous hydrogels, “cryogels,” are produced by polymerization in a partially frozen state when the ice crystals perform as a porogen. Cryogels have a unique combination of properties: (i) large (10–100 μm) pores; (ii) minimal non-specific interactions due to the hydroph ...

Methods are presented for the preparation, ligand density analysis and use of an affinity adsorbent for the purification of a glutathione S-transferase (GST) fusion protein in packed and expanded bed chromatographic processes. The protein is composed of GST fused to a zinc finger transcr ...

Enzymes immobilized on solid-phase matrices have found various applications in biotechnology, molecular biology and molecular diagnostics and can serve as industrial catalysts and as specific reagents for analytical procedures. A wide range of supports have been utilized for i ...

The interaction between avidin and biotin or streptavidin and biotin forms the basis of several widely used immunohistochemical techniques. An assumption inherent to these techniques is that endogenous biotin is not present in the tissue in detectable quantities, as neither avidin n ...

Immunodetection techniques are dependent on enzyme–protein conjugates for the visualization of antigen–antibody complexes. One of the most widely used is the avidin–biotin–peroxidase complex (ABC) method. However, treatment of certain tissues with ABC reagents alone may res ...

Posttranslational modifications of histones play an important role in the regulation of chromatin structure and, hence, gene regulation. Recently, we have identified a novel modification of histones: binding of the vitamin biotin to lysine residues in histones H2A, H3, and H4. Here, we des ...